Abstract
ColE1 DNA has been allowed to react in vitro with N-acetoxy-N-2-[14C]acetylaminofluorene in the range of 0-15 N-2-[14C]acetylaminofluorene residues bound per molecule of DNA, at the C8 of guanine residues. Purified rabbit antibodies to both N-2-(guanosine-8-yl)-acetylaminofluorene and native DNA that had reacted with N-acetoxy-N-2-acetylaminofluorene were shown by electron microscopy to recognize specifically the acetylaminofluorene-modified ColE1 DNA. The antibodies bound to DNA were visualized either per se or after reaction with goat anti-rabbit immunoglobulins coupled with ferritin. There was a linear relationship between the average number of antibodies bound per DNA molecule and the number of N-2-(deoxyguanosine-8yl)-acetylaminofluorene residues per DNA molecule. The slope of this straight line was equal to 0.4. Due to the bivalence of the immunoglobulins one would expect a value of 0.5; we actually observed an important fraction of the bound antibodies crosslinking two parts of the same (or of another) DNA molecule.
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- Clewell D. B. Nature of Col E 1 plasmid replication in Escherichia coli in the presence of the chloramphenicol. J Bacteriol. 1972 May;110(2):667–676. doi: 10.1128/jb.110.2.667-676.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dubochet J., Ducommun M., Zollinger M., Kellenberger E. A new preparation method for dark-field electron microscopy of biomacromolecules. J Ultrastruct Res. 1971 Apr;35(1):147–167. doi: 10.1016/s0022-5320(71)80148-x. [DOI] [PubMed] [Google Scholar]
- Fuchs R. P. Arylamidation and arylation by the carcinogen N-2-fluorenylacetamide: a sensitive and rapid radiochemical assay. Anal Biochem. 1978 Dec;91(2):663–673. doi: 10.1016/0003-2697(78)90553-5. [DOI] [PubMed] [Google Scholar]
- Guigues M., Leng M. Reactivity of antibodies to guanosine modified by the carcinogen N-acetoxy-N-2-acetylaminofluorene. Nucleic Acids Res. 1979 Feb;6(2):733–744. doi: 10.1093/nar/6.2.733. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kriek E. Persistent binding of a new reaction product of the carcinogen N-hydroxy-N-2-acetylaminofluorene with guanine in rat liver DNA in vivo. Cancer Res. 1972 Oct;32(10):2042–2048. [PubMed] [Google Scholar]
- Lefèvre J. F., Fuchs R. P., Daune M. P. Comparative studies on the 7-iodo and 7-fluoro derivatives of N-acetoxy-N-2-acetylaminofluorene: binding sites on DNA and conformational change of modified deoxytrinucleotides. Biochemistry. 1978 Jun 27;17(13):2561–2567. doi: 10.1021/bi00606a016. [DOI] [PubMed] [Google Scholar]
- Leng M., Sage E., Fuchs R. P., Duane M. P. Antibodies to DNA modified by the carcinogen N-acetoxy-N-2-acetylaminofluorene. FEBS Lett. 1978 Aug 15;92(2):207–210. doi: 10.1016/0014-5793(78)80755-8. [DOI] [PubMed] [Google Scholar]
- Poirier M. C., Dubin M. A., Yuspa S. H. Formation and removal of specific acetylaminofluorene-DNA adducts in mouse and human cells measured by radioimmunoassay. Cancer Res. 1979 Apr;39(4):1377–1381. [PubMed] [Google Scholar]
- Poirier M. C., Yuspa S. H., Weinstein I. B., Blobstein S. Detection of carcinogen-DNA adducts by radiommunoassay. Nature. 1977 Nov 10;270(5633):186–188. doi: 10.1038/270186a0. [DOI] [PubMed] [Google Scholar]
- Sage E., Fuchs R. P., Leng M. Reactivity of the antibodies to DNA modified by the carcinogen N-acetoxy-N-acetyl-2-aminofluorene. Biochemistry. 1979 Apr 3;18(7):1328–1332. doi: 10.1021/bi00574a032. [DOI] [PubMed] [Google Scholar]
- Tomizawa J., Sakakibara Y., Kakefuda T. Replication of colicin E1 plasmid DNA in cell extracts. Origin and direction of replication. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2260–2264. doi: 10.1073/pnas.71.6.2260. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Voordouw G., Kam Z., Borochov N., Eisenberg H. Isolation and physical studies of the intact supercoiled, the open circular and the linear forms of ColE1-plasmid DNA. Biophys Chem. 1978 May;8(2):171–189. doi: 10.1016/0301-4622(78)80008-8. [DOI] [PubMed] [Google Scholar]
- Westra J. G., Kriek E., Hittenhausen H. Identification of the persistently bound form of the carcinogen N-acetyl-2-aminofluorene to rat liver DNA in vivo. Chem Biol Interact. 1976 Oct 2;15(2):149–164. doi: 10.1016/0009-2797(76)90160-5. [DOI] [PubMed] [Google Scholar]
- de Murcia G., Das G. C., Erard M., Daune M. Superstructure and CD spectrum as probes of chromatin integrity. Nucleic Acids Res. 1978 Feb;5(2):523–535. doi: 10.1093/nar/5.2.523. [DOI] [PMC free article] [PubMed] [Google Scholar]